Electrophoretic Analysis of Human Parotid Salivary Proteins with Application to the Study of Rheumatoid Arthritis and Sjogren's Syndrome

Human parotid saliva contains many proteins with diverse functions. In the course of a number of diseases, especially where the normal function of the salivary gland is affected, changes may occur in the levels of certain of these. Therefore the analysis of some of these proteins may be of diagnostic significance. This study has focused on the development and refinement of electrophoretic and of protein detection techniques in order to allow the fractionation of proteins in small volumes of human saliva with the minimum of sample preparation. In order to give an example of their possible diagnostic significance, the electrophoretic techniques which were developed were applied to the fractionation and partial characterisation of the anionic salivary proteins associated with connective tissue disorders such as rheumatoid arthritis and Sjogren's Syndrome. The saliva of patients with rheumatoid arthritis and Sjogren's Syndrome contains additional anionic proteins, which are either present in very low levels or below detection limits in the saliva of normal healthy individuals. Research into the identity of these proteins has been largely hindered by the relatively high electrolyte and low protein content of human parotid saliva, making it necessary to desalt and concentrate the saliva samples prior to carrier ampholyte-based isoelectric focusing. Desalting requires relatively large volumes (preferably > 2ml) of saliva, which may be difficult or even impossible to obtain from diseased glands. Also, one-dimensional isoelectric focusing cannot separate these anionic proteins from the acidic proline-rich proteins of human saliva, as both groups of proteins have overlapping isoelectric points. In this study, a hybrid carrier ampholyte-immobilised pH gradient isoelectric focusing technique was developed to analyse human salivary proteins. Immobilised pH gradients (IPG's) of 3 pH ranges were prepared: broad-range (pH 4-9) IPG was used for the general study of human salivary proteins; while 2 narrow, acidic range IPG's (pH 2.8-4.5 and pH 3.5-5.0) were used to analyse proteins of low isoelectric points, such as the anionic salivary proteins associated with rheumatoid arthritis and Sjogren's Syndrome. This method allowed the difficulties involved when conventional carrier ampholyte-based isoelectric focusing is used to be circumvented, thus making it possible to fractionate the proteins in small volumes (approximately 50ul) of human saliva without prior treatment except for centrifugation. Parotid salivary proteins were also analysed by onedimensional SDS-PAGE. SDS-PAGE gels were subjected to im-muno- and lectin affinity-blotting in order to characterise or identify some of the protein bands. Proline-rich proteins were recognised by their characteristic pink-staining with the dye Coommassie Brilliant Blue R250, and some of the bands which were revealed have been correlated with proline-rich proteins which have been isolated and partially characterised by other research groups. SDS-PAGE failed to reveal any obvious differences between the band patterns of normal subjects and those of patients with rheumatoid arthritis or Sjogren's Syndrome. Two-dimensional gel electrophoresis was also carried out using hybrid carrier ampholyte-immobilised pH gradient polyacrylamide gels in the first dimension and thin-layer SDS-polyacrylamide gradient gels in the second. By means of a combination of staining and electroblotting of the two-dimensional gels onto nitrocellulose followed by probing with specific antisera, a two-dimensional map of human parotid salivary proteins, in which most of the major components have been identified, has been obtained. These techniques were applied to the investigation of the nature of the anionic salivary proteins associated with rheumatoid arthritis and Sjogren's Syndrome. Two-dimensional electrophoresis with pH 3.5-5.0 IPG's in the first dimension followed by silver staining revealed these proteins to be heterogeneous (pI's approximately 3.65-4.75) and of a single relative molecular weight (approximately 32,000). In normal healthy controls these silver stained components were less heterogeneous (pI's approximately 3.65-4.25). Incubation with neuraminidase showed that their heterogeneity was largely due to differing contents of sialic acid in their carbohydrate side-chains. In order to attempt to identify these proteins, the one-dimensional IPG and two-dimensional gels were electroblotted and the blots were probed with a variety of antisera. The proteins appeared to be immunoreactive with antisera to human tissue kallikrein, a protein the level of which has often been reported to be elevated in the saliva of patients with Sjogren's Syndrome. (Abstract shortened by ProQuest.)

Penentuan Hasil Tindakbalas Antara Asid Kromotropik Dan Formaldehid

Identiti tindakbalas antara asid kromotropik dan formaldehid berlebihan tanpa asid sulfuric pekat dibuktikan sebagai dimer di mana dua molekul asid kromotropik diikat melalui dua titisan olefinik, berasaskan kepada spectrum N.M.R Proton, mikroanalisisuntuk C,H, N dan S, dan pentitratan Karl Fischer

Investigation of twenty selected medicinal plants from Malaysia for anti-Chikungunya virus activity

Chikungunya virus is a reemerging arbovirus transmitted mainly by Aedes mosquitoes. As there are no specific treatments available, Chikungunya virus infection is a significant public health problem. This study investigated 120 extracts from selected medicinal plants for anti-Chikungunya virus activity. The plant materials were subjected to sequential solvent extraction to obtain six different extracts for each plant. The cytotoxicity and antiviral activity of each extract were examined using African monkey kidney epithelial (Vero) cells. The ethanol, methanol and chloroform extracts of Tradescantia spathacea (Commelinaceae) leaves showed the strongest cytopathic effect inhibition on Vero cells, resulting in cell viabilities of 92.6% ± 1.0% (512 μg/ml), 91.5% ± 1.7% (512 μg/ml) and 88.8% ± 2.4% (80 μg/ml) respectively. However, quantitative RT-PCR analysis revealed that the chloroform extract of Rhapis excelsa (Arecaceae) leaves resulted in the highest percentage of reduction of viral load (98.1%), followed by the ethyl acetate extract of Vernonia amygdalina (Compositae) leaves (95.5%). The corresponding 50% effective concentrations (EC50) and selectivity indices for these two extracts were 29.9 ± 0.9 and 32.4 ± 1.3 μg/ml, and 5.4 and 5.1 respectively. Rhapis excelsa and Vernonia amygdalina could be sources of anti-Chikungunya virus agents. [Int Microbiol 19(3):175-182 (2016)]Keywords: Chikungunya virus · antivirals · cytotoxicity · sequential extraction · medicinal plant

Characterisation of Bidor Kaolinite and Illite

Two types of commercial clay minerals from the Bidor region of Perak were studied. From the X-ray diffraction study, these clays were identified as disordered kaolinite and 2 M polymorph of illite. The morphologies, the surface properties, the pore structures and the infrared absorption properties were examined. The basis for the identification of these clays is discussed

A Storm in a "T" Cup

We revisit the process of transversification and agglomeration of particle momenta that are often performed in analyses at hadron colliders, and show that many of the existing mass-measurement variables proposed for hadron colliders are far more closely related to each other than is widely appreciated, and indeed can all be viewed as a common mass bound specialized for a variety of purposes.Comment: 3 pages, 2 figures, presented by K.C. Kong at the 19th Particles and Nuclei International Conference, PANIC 2011, MIT, Cambridge, MA (July 24-29, 2011

Interactions between Plant Extracts and Cell Viability Indicators during Cytotoxicity Testing: Implications for Ethnopharmacological Studies

Purpose: To compare the cytotoxicity of six medicinal plants (Acmella ciliata, Amaranthus tricolor, Coriandrum sativum, Glebionis coronaria, Kyllinga brevifolia and Tradescantia zebrina) using 3-(4, 5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red uptake (NRU) assays.Methods: Hexane, chloroform, ethyl acetate, ethanol, methanol and water extracts were obtained for each plant by sequential solvent extraction. Cytotoxicity was evaluated in triplicate, from 640 to 5 μg/mL, two-fold, serially on monkey kidney epithelial (Vero) cells.Results: The hexane, chloroform and ethyl acetate extracts of the six plants were more toxic to the Vero cells compared to the ethanol, methanol and water extracts. Thirty one percent (11/36) and 75 % (27/36) of the extracts showed significant cytotoxicity (p < 0.05) in MTT and NRU assays, respectively. The 78, 52 and 7 % cytotoxicity levels detected in 27 extracts using the MTT assay were significantly (p < 0.05) underestimated at 640, 320 and 160 μg/mL, respectively, using NRU assay. Nine extracts from five plants exhibited significantly lower (p < 0.05) 50 % cytotoxic concentration (CC50) when NRU assay was employed compared to MTT assay. At 640 μg/mL, 10 of the 21 extracts were also found to react chemically with MTT, causing a 2.0 – 29.1-fold increase in the absorbance value (550 nm) compared to control.Conclusion: The plant extracts of A. ciliata, A. tricolor, C. sativum, G. coronaria, K. brevifolia and T. zebrina show concentration- and extraction method-dependent cytotoxicity using MTT and NRU assays. NRU assay appears to be more sensitive and reliable than MTT assay for cell viability evaluation of the plant extracts.Keywords: Acmella ciliata, Amaranthus tricolor, Coriandrum sativum, Glebionis coronaria, Kyllinga brevifolia and Tradescantia zebrina, Extraction, Medicinal plant, Neutral red uptake assay, Vero cel

Citrus Bent Leaf Viroid

Citrus bent leaf viroid (CBLVd) from genus Apscaviroid, is one of the widely distributed viroids among the seven citrus viroids. It is comprised of three variants: Citrus viroid-Ia (CVd-Ia) (327 - 329 nucleotides), Citrus viroid-Ib (CVd-Ib) (315 - 319 nucleotides) and Citrus viroid-I-low sequence similarity (CVd-I-LSS) (325 - 330 nucleotides). Virulence of CBLVd totally expressed on citrus plants. Etrog citron (Citrus medica (L.)) coinfected with CBLVd, Citrus exocortis viroid (CEVd), Citrus viroid-III (CVd-III) and Citrus viroid-V (CVd-V) showed epinasty, leaf rolling, and stunting. CBLVd has been reported to reduce the canopy proportion and fruit production of citrus trees inserted on trifoliate orange rootstock. Moreover, citrus tree infected with singly CBLVd or in combinations with CEVd, Hop stunt viroid (CVd-II) and CVd-III induced dwarfing have been associated with poor development of the root system. Reverse-transcriptase polymerase chain reaction (RT-PCR) amplification and multiplex reverse-transcriptase polymerase chain reaction (MRT-PCR) amplification have been widely used to detect citrus viroids including CBLVd. As citrus viroids are emerging threats in citrus groves, therefore, this review covers the evolution, geographical distribution and epidemiology, economic impact and symptomatology, host range and transmission, detection, and management will be helpful in formulating the integrated management strategies for CBLVd

A quantum tug of war between randomness and symmetries on homogeneous spaces

We explore the interplay between symmetry and randomness in quantum information. Adopting a geometric approach, we consider states as $H$-equivalent if related by a symmetry transformation characterized by the group $H$. We then introduce the Haar measure on the homogeneous space $\mathbb{U}/H$, characterizing true randomness for $H$-equivalent systems. While this mathematical machinery is well-studied by mathematicians, it has seen limited application in quantum information: we believe our work to be the first instance of utilizing homogeneous spaces to characterize symmetry in quantum information. This is followed by a discussion of approximations of true randomness, commencing with $t$-wise independent approximations and defining $t$-designs on $\mathbb{U}/H$ and $H$-equivalent states. Transitioning further, we explore pseudorandomness, defining pseudorandom unitaries and states within homogeneous spaces. Finally, as a practical demonstration of our findings, we study the expressibility of quantum machine learning ansatze in homogeneous spaces. Our work provides a fresh perspective on the relationship between randomness and symmetry in the quantum world.Comment: 9 + 1 pages, 3 figure

Total phenolic and antioxidant activities of Pouteria campechiana fruit parts

This study aimed to evaluate the total phenolics and antioxidant capacities of the seeds, pulp and peel of Pouteria campechiana fruit using three extraction solvents (water, 70% methanol and 70% ethanol). Among them, 70% ethanol exhibited the best solvent for yielding highest total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activities. The results showed that 70% ethanol extract from the peel contained the highest TPC(2304.7 mg gallic acid equivalent/100 g dw) while the pulp has the highest TFC(6414.03 mg rutin equivalent/100 g dw). The antioxidant activities of the pulp and peel ethanolic extracts were high as determined using 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation assay (49.60 and 49.56 mmoL TE/100 g dw) and ferric reducing antioxidant power assay (43.88 and 42.94 Fe2+/100 g dw) but not for seeds. However, their diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities were ~88%. Thus, the pulp and peel of P. campechiana fruit can be utilized as natural source for antioxidant components

Identification of material properties of orthotropic composite plate using hybrid non-destructive evaluation approach

Identification of material properties is one of the key issues in composite materials research. The mechanical properties of composite materials depend on diverse factors such as configuration of the laminates, constituent materials used and production method adopted. Conventional testing approach tends to be time-consuming, expensive and destructive. As an alternative, a rapid, inexpensive, hybrid and non-destructive evaluation approach which utilises experimental modal analysis and finite element analysis is proposed. Experimental modal data which consist of natural frequencies and mode shapes of an orthotropic composite plate are utilised for correlation purpose with its finite element model. This finite element model of the composite plate is continuously updated and achieves less than 5% in difference of natural frequencies and over 70% in modal assurance criterion. Material properties such as Young's moduli, inplane shear modulus and Poisson ratio of the composite plate are then successfully determined using the well-correlated FE model